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Insulin Self-association: Effects on Lung Disposition Kinetics in the Airways of the Isolated Perfused Rat Lung (IPRL)

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Abstract

Purpose

To characterize the kinetic dependence of pulmonary absorption and metabolism of insulin and lispro on the magnitude of their hexameric association.

Methods

Hexamer content by weight percent (%Hex) in various insulin-zinc and lispro-zinc solutions were determined by quantitative centrifugal ultrafiltration and zinc titration with terpyridine (QCUF-ZTT). Each of the solutions (0.1 ml) was then administered into the airways of the IPRL of normal and experimental diabetic animals. Rate constants were determined for lung absorption (k a) and non-absorptive loss (k nal; comprising mucociliary clearance and metabolism).

Results

%Hex in administered solutions ranged from 3.3 to 94.4%. Data analysis showed excellent correlations between the values for k a or k nal and %Hex, irrespective of insulin type, concentration, solution pH or ionic strength. The values for k a decreased (0.22 → 0.05 h−1) with increasing %Hex, as did values for k nal. At %Hex in administered solutions ≥50%, values for k nal approached estimates for the rate constant for mucociliary clearance, implying that lung metabolism occurred primarily with monomeric insulin. There were no differences in insulin disposition kinetics between lungs taken from experimental diabetic and sham-control animals.

Conclusions

The kinetics of pulmonary insulin disposition depended on the magnitude of molecular self-association. Dissociated forms of insulin (dimers or monomers) in the dosing solution showed higher rates than hexamers for both lung absorption and metabolism.

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Abbreviations

BSA:

bovine serum albumin

CBS:

citrate-buffered saline

CD:

circular dichroism

COD:

coefficient of determination

ELISA:

enzyme-linked immunosorbent assay

%Hex:

percent of hexamers by weight

IPRL:

isolated perfused rat lung

IU:

International Unit

LLF:

lung lining fluid

MSC:

model selection criterion

NIH:

National Institute of Health

PBS:

phosphate-buffered saline

QCUF–ZTT:

quantitative centrifugal ultrafiltration and zinc titration with terpyridine

%RSD:

percent of relative standard deviation

SD:

standard deviation

SE:

standard error

STZ:

streptozotocin

t 1/2 :

apparent half-life

USP:

United States Pharmacopeia

UV:

ultraviolet

VCU:

Virginia Commonwealth University

μ :

ionic strength

[θ]:

mean residue molar ellipticity

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Acknowledgments

The authors are grateful to Eli Lilly and Company for their gift of lispro-zinc (Lispro-Zn), and to Verne G. Schirch, Ph.D. (Department of Biochemistry, VCU) for the use and assistance with CD spectrometry. The Medical College of Virginia Foundation and VCU’s A.D. Williams Research Funds (MS) supported this research. YP acknowledges VCU School of Pharmacy financial support during her graduate study.

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Author notes

  1. Yinuo Pang

    Present address: Schering-Plough Biopharma, Palo Alto, California, USA

Authors and Affiliations

  1. Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, 410 N. 12th Street, P.O. Box 980533, Richmond, Virginia, 23298-0533, USA

    Yinuo Pang, Masahiro Sakagami & Peter R. Byron

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  1. Yinuo Pang
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  2. Masahiro Sakagami
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  3. Peter R. Byron
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Correspondence to Masahiro Sakagami.

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Pang, Y., Sakagami, M. & Byron, P.R. Insulin Self-association: Effects on Lung Disposition Kinetics in the Airways of the Isolated Perfused Rat Lung (IPRL). Pharm Res 24, 1636–1644 (2007). https://doi.org/10.1007/s11095-007-9292-6

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